Masters M, Paterson T, Popplewell A G, Owen-Hughes T, Pringle J H, Begg K J
Department of Molecular Biology, University of Edinburgh, Scotland.
Mol Gen Genet. 1989 Apr;216(2-3):475-83. doi: 10.1007/BF00334393.
The DnaA protein of Escherichia coli, essential for initiation at oriC, binds at a defined sequence which occurs at the chromosomal origin, near plasmid replication origins and in the promoters of the dnaA and mioC genes. This sequence also occurs at many other sites on the E. coli chromosome including three sites within the essential cell division genes ftsQ and A. Using an fts-lac fusion phage, lambda JFL100, we show here that fts gene expression responds both to reduced and increased intracellular levels of DnaA protein in a manner consistent with the hypothesis that DnaA protein regulates fts gene expression. Experiments using dnaC and dnaB-ts strains, however, suggest that DnaA control of fts transcription may be indirect, at least in part, with fts responding to the rate of initiation at oriC as well as to changes in DnaA protein level per se. It differs in this respect from dnaA gene expression which is unaffected when initiation of replication is inhibited by DnaB or DnaC inactivation. Strains integratively suppressed with pKN500 behave anomalously; neither fts nor dnaA transcription is significantly increased when DnaA is inactivated in these strains.
大肠杆菌的DnaA蛋白对于在oriC起始复制至关重要,它结合在特定序列上,该序列出现在染色体起点、质粒复制起点附近以及dnaA和mioC基因的启动子中。此序列在大肠杆菌染色体的许多其他位点也有出现,包括必需的细胞分裂基因ftsQ和A内的三个位点。利用fts - lac融合噬菌体λJFL100,我们在此表明fts基因表达对细胞内DnaA蛋白水平的降低和升高均有反应,这与DnaA蛋白调节fts基因表达的假说一致。然而,使用dnaC和dnaB - ts菌株进行的实验表明,DnaA对fts转录的控制可能至少部分是间接的,fts对oriC处的起始速率以及DnaA蛋白水平本身的变化均有反应。在这方面它与dnaA基因表达不同,当复制起始因DnaB或DnaC失活而受到抑制时,dnaA基因表达不受影响。用pKN500进行整合抑制的菌株表现异常;当这些菌株中的DnaA失活时,fts和dnaA转录均未显著增加。